Objective To explore the possibilityof constructing tissue engineering muscles by combining allogeneic myoblasts with small instestinal submucosa(SIS) in rabbits.Methods A large number of purified myoblasts were obtained with multiprocedure digestion and repeated attachment method from skeletal muscles taken from extremities of immature rabbits which were born 7 days ago. The myoblasts were labeled with BrdU, and then combined with SIS to construct tissue engineering muscles. This kind of tissue engineering muscles were grafted into the gastrocnemius muscle defect (1.5 cm in length, 1.0 cmin width) of fifteen rabbits as the experimental group. The SIS was grafted into the same position in the control group. The rabbits were sacrificed 4, 6, 8 weeks after operation. The tissue engineering muscles were evaluated by macroscopic、histological and immunohistochemical observations, and by quantitative analysis of local immunocyte in the grafting site. Results Allogeneic myoblasts with SIS were combined perfectly in vitro. The SIS was connected tightly to surrounding skeletal muscles and inflammation response was obvious 4 weeks after grafting.The SIS began to break down and inflammation response became slight 6 and 8 weeks after operation. Compared with that of 8th week, the quantitative analysis oflocal immunocyte in 4th and 6th week in both experimental and control group hassignificance(Plt;0.05). Newly formed muscle tissues were found around SIS in the experimental group in 4th, 6th, and 8th week. Expression of BrdU and myosin immunohistochemical staining were positive in the experimental group and negative inthe control group.Conclusion Tissue engineering muscles of rabbits which are constructed by combining allogeneic myoblasts with SIS can survive and proliferate.
In order to investigate the possibility of repairing injuried tendon with living artificial tendon, after combining culture, subcultured autogenous tendon cells with carbon fibers were implanted into the calcaneous tendon of rabbits. In different stages, the synthesis of type I collagen and their relevant morphological changes were observed. The results showed as follows: after implantation, tendon cells continued proliferating. Four weeks after implantation, tendon cells were detached from the carbon fibers and proliferated and produced collagen among the carbon fibers. The collagen fibrils were linked with each other to formed a dense structure. In the linkage site, the collagen fibrils originated from the implants joined to that from the ruptured end of the tendon, which meaned that the implant was healed with the recipient tendon. Observed under scanning electronic microscope, the tendon cells were lined among the carbon fibers evenly and in order, the collagen fibrils joined each other and formed an network, the fibrils were lined parallel to the carbon fibers. Under transparent electron microscope, the nucleolus were clear and organelle were abundant.